Imidazole derivatives or their salts

ABSTRACT

Imidazole derivatives of the formula (1) or salts thereof, pharmaceuticals containing the derivatives or the salts, and intermediates for the synthesis of the derivatives or the salts (wherein R 1  is lower alkyl; R 2  is alkyl or aralkyl; and X 1  is halogeno). These compounds exhibit G-CSF-like activities and can be substituted for G-CSF preparations.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a national-stage filing under 35 U.S.C. 371 ofPCT/JP01/08231, filed Sep. 21, 2001. This application also claimspriority to JAPAN 2000-288589, filed Sep. 22, 2000 and JAPAN2000-394273, filed Dec. 26, 2000.

TECHNICAL FIELD

The present invention relates to novel imidazole derivatives exhibitingcytokine-like activities; in particular, activities analogous to thoseexhibited by granulocyte-colony stimulating factor (hereinafter referredto as “G-CSF”).

BACKGROUND ART

G-CSF drugs have been found effective in therapy of immune diseaseswhich accompany reduction in neutrophils caused by chemotherapy orradiotherapy of cancer. In clinical settings, a human gene recombinantG-CSF injection (Sankyo Co., Ltd.) or similar products are currentlyemployed as G-CSF drugs. However, the G-CSF injection involvesdrawbacks; namely, it produces problematic side effects, such asinflammation, bone pain, fever, and chill, and upon oral administrationit is rapidly degraded, because it is composed of glycoproteins.Therefore, in order to solve these problems, there remains need fordevelopment of a low-molecular weight compound which exhibits G-CSF-likeactivity and which can serve as a substitute for G-CSF drugs. However,presently, very few compounds, such as those described in WO97/44033,are known to exhibit G-CSF-like activities.

Accordingly, an object of the present invention is to provide a novellow-molecular-weight compound exhibiting cytokine-like activities,particularly G-CSF-like activities.

Numerous studies have been carried out on imidazole compounds forpossible application as agrochemicals or pharmaceuticals. Of theimidazole compounds, those having a 2-alkoxyphenyl group are reported inJapanese Patent Application Laid-Open (kokai) Nos. 63-146864 or61-267557. However, these publications fail to mention whether or notthose compounds exhibit cytokine-like activities. Moreover, a2-alkoxyphenyl derivative having a halogen group at the 5-position ofthe phenyl group has not yet been reported.

DISCLOSURE OF THE INVENTION

Under the above circumstances, the present inventors performed screeningtests on a variety of compounds, in search of low-molecular-weightcompound having G-CSF-like activities, and have found that a novel classof imidazole derivatives (1) having a 2-alkoxy-5-halogenophenyl groupexhibits cytokine-like activities; in particular, G-CSF-like activities,thus leading to completion of the invention.

The present invention provides an imidazole derivative represented bythe following formula (1):

(wherein R¹ represents a lower alkyl group, R² represents an alkyl oraralkyl group, and X¹ represents a halogen atom) or a salt thereof, andan intermediate for the synthesis of the derivative or the salt.

The present invention also provides a drug containing an imidazolederivative of formula (1) or a salt thereof as an active ingredient.

The present invention also provides a pharmaceutical compositioncontaining an imidazole derivative of formula (1) or a salt thereof, anda pharmacologically acceptable carrier.

The present invention also provides use of an imidazole derivative offormula (1) or a salt thereof in manufacture of a drug.

The present invention further provides a method of treating an immunedisease accompanying reduction in neutrophils, the method comprisingadministration of an imidazole derivative of formula (1) to a patient inneed thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the multiplication ability of human G-CSF-dependent cellsin the presence of rhG-CSF; and

FIG. 2 shows induction of multiplication of human G-CSF-dependent cellsby the compound of the present invention.

BEST MODE FOR CARRYING OUT THE INVENTION

In formula (1), lower alkyl groups represented by R¹ include C1-C5linear, branched, or cyclic alkyl groups, and specific examples thereofinclude methyl, ethyl, n-propyl, n-butyl, n-pentyl, and cyclopropyl.

Examples of the alkyl group represented by R² include C3-C15 linear orbranched alkyl groups, and specific examples thereof include n-butyl,n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, andn-dodecyl.

Examples of the aralkyl group represented by R² include phenyl C1-C5alkyl groups such as benzyl and phenethyl, and halo-substituted groupsthereof, and specific examples thereof include benzyl, 4-chlorobenzyl,2,4-dichlorobenzyl, 2-fluorobenzyl, phenethyl, and 2-fluorophenethyl.

Examples of the halogen represented by X¹ include fluorine, chlorine,bromine, and iodine, with fluorine and chlorine being preferred.

No particular limitation is imposed on the salts of the imidazolederivatives (1) of the present invention, so long as they arepharmacologically acceptable salts. Examples of the salts include, butare not limited to, hydrochloride, nitrate, hydrobromide,p-toluenesulfonate, methanesulfonate, fumarate, succinate, and lactate.

The imidazole derivatives (1) or their salts of the present inventionhave optical isomers based on asymmetric carbon atoms, and opticallyactive compounds and mixtures of isomers such as racemic modificationsare encompassed by the present invention. In addition, hydrates of thesecompounds are also encompassed by the present invention.

The imidazole derivatives (1) or their salts of the present inventionmay be produced in accordance with the following reaction formula.

[wherein R¹, R², and X¹ have the same meanings as described above, andX² denotes halogen].

Briefly, a known compound (3) is halogenized, and the resultanthaloketone compound (4) is transformed into an imidazole derivative. Theobtained compound (5) is subjected to a Grignard reaction, to therebyyield a compound (2), which is finally alkylated or aralkylated,yielding an imidazole derivative (1) of the present invention. In thisconnection, the compound (2) also is a novel compound, and is useful asan intermediate for the synthesis of imidazole derivative (1).Hereinafter, the synthesis method of the present invention is describedin accordance with the above steps.

(Step A)

A halogenizing agent is reacted with compound (3), to thereby yieldhaloketone compound (4).

Of a variety of compounds (3), examples of preferred class of compoundsin which X¹ is chlorine or fluorine include those commercially availablefrom Aldorich. Examples of X² in compound (4) include chlorine, bromine,iodine, with chlorine and bromine being preferred. Preferably, thehalogenizing agent is cupric bromide, bromine, sulfuryl chloride, or asimilar substance. As the reaction solvent, an ethyl acetate-chloroformsolvent mixture, 1,4-dioxane, diethyl ether, a 1,4-dioxane-diethyl ethersolvent mixture, acetic acid, dichloromethane, or a similar substancemay be used.

(Step B)

Compound (4) is reacted with imidazole, to thereby yield compound (5).

Preferably, the amount of the imidazole to be employed is 2 to 5equivalents with respect to compound (4). As the reaction solvent,N,N-dimethylformamide, dimethyl sulfoxide, methanol, ethanol,tetrahydrofuran, dioxane, or a similar substance may be used singly oras a mixture, with N,N-dimethylformamide being particularly preferred.Preferably, the reaction temperature is 0 to 50° C., and the reactiontime is 1 to 200 hours, more preferably 5 to 48 hours.

(Step C)

Compound (5) is reacted with a Grignard reagent, to thereby attainsynthesis of compound (2).

As the Grignard reagent, a compound having the aforementioned loweralkyl group R¹ may be used, and examples of the Grignard reagent includemethylmagnesium chloride, methylmagnesium bromide, methylmagnesiumiodide, ethylmagnesium chloride, ethylmagnesium bromide, propylmagnesiumchloride, propylmagnesium bromide, cyclopropylmagnesium chloride, andcyclopropylmagnesium bromide. Preferably, the amount of the Grignardreagent to be employed is 1 to 50 equivalents with respect to compound(5). Preferably, the reaction solvent is tetrahydrofuran, diethyl ether,or a similar substance. Preferably, the reaction temperature is 0 to 50°C., and the reaction time is 1 to 200 hours, more preferably 5 to 48hours.

(Step D)

Compound (2) is reacted with alkyl halide or aralkyl halide in thepresence of a base, to thereby yield compound (1) of the presentinvention.

As the alkyl halide or aralkyl halide, a halide having theaforementioned R² as an alkyl group or aralkyl group may be used.Examples of halogens of the halide include chlorine, bromine, andiodine, with bromine being particularly preferred. The amount of thealkyl halide or aralkyl halide to be employed is preferably 1 to 50,more preferably 1 to 10 equivalents with respect to compound (2).Examples of bases include sodium hydroxide, potassium hydroxide, bariumhydroxide, sodium carbonate, potassium carbonate, sodium hydride,potassium hydride, sodium methoxide, sodium ethoxide, pyridine, andtriethylamine, with sodium hydroxide and potassium hydroxide beingparticularly preferred. The amount of the base to be employed ispreferably 1 to 50, more preferably 1 to 10 equivalents with respect tocompound (2). Any reaction solvents may be employed so long as they donot react with the starting compound. For example, there may be employedamides such as N,N-dimethylformamide and N,N-dimethylacetamide; alcoholssuch as methanol, ethanol, 1-propanol, 2-propanol, 1-butanol,2-methyl-2-propanol, ethylene glycol, propylene glycol, glycerin, andmethyl cellosolve; ethers such as tetrahydrofuran, dioxane, anddimethoxyethane; and dimethyl sulfoxide. These may be used singly or asmixtures. Of these N,N-dimethylformamide is particularly preferred. Thereaction temperature is preferably 0 to 100° C., more preferably 10 to50° C. The reaction time is preferably 1 to 200 hours, more preferably 5to 48 hours.

The imidazole derivatives (1) or their salts of the present inventioncan also be produced in accordance with the following reaction formula:

[wherein R¹, R² and X¹ have the same meanings as described above].

Briefly, a known compound (6) is acylated, and the resultant compound(7) is subjected to a Fries rearrangement reaction for conversion intocompound (8), which is then alkylated or aralkylated to yield compound(9). The compound (9) is epoxidized into a compound (10) and thenreacted with an imidazole, to thereby obtain the imidazole derivative(1) of the present invention. The present synthesis method will next bedescribed in accordance with the above steps.

(Step E)

Compound (6) is acylated with acid anhydride, or with acid halide in thepresence of a base, to thereby synthesize compound (7).

In the former case, the acid anhydride may be a compound having theaforementioned lower alkyl group R¹. Examples of the acid anhydrideinclude acetic anhydride, propionic anhydride, butyric anhydride, andvaleric anhydride. The amount of the acid anhydride to be used ispreferably 1 to 50, more preferably 1 to 5 equivalents with respect tocompound (6). A solvent free system is preferred, but a solvent may beadded. Also, a catalytic amount of an acid is desirably added. Examplesof the acids include hydrochloric acid, sulfuric acid, and nitric acid,with sulfuric acid being particularly preferred. The reactiontemperature is preferably 0 to 100° C., more preferably 10 to 50° C. Thereaction time is preferably 1 to 200 hours, more preferably 5 to 48hours.

In the latter case, the acid halide may be a compound having theaforementioned lower alkyl group R¹. For example, mention may be givenof acetyl chloride, acetyl bromide, propionyl chloride, propionylbromide, butyryl chloride, valeryl chloride, and cyclopropanecarbonylchloride. The amount of the acid halide to be employed is preferably 1to 50, more preferably 1 to 5 equivalents with respect to compound (6).Preferably, the reaction solvent is dichloromethane or a similarsubstance. Preferably, the base is pyridine, triethylamine, or a similarsubstance. The amount of the base to be employed is preferably 1 to 50,more preferably 1 to 5 equivalents with respect to compound (6). Thereaction temperature is preferably −20 to 100° C., more preferably 0 to50° C. The reaction time is preferably 1 to 200 hours, more preferably 5to 48 hours.

(Step F)

Compound (7) is heated in the presence of a Lewis acid, in the presenceor absence of a reaction solvent, to thereby synthesize compound (8).

Examples of the Lewis acid include aluminum chloride, aluminum bromide,zinc chloride, zinc bromide, and stannic chloride, with aluminumchloride being particularly preferred. The amount of the Lewis acid tobe employed is preferably 1 to 50, more preferably 1 to 5 equivalentswith respect to compound (7). Preferably, the reaction solvent isdichloromethane or a similar substance. Solvent-free systems are alsopreferred. The reaction temperature is preferably −20 to 200° C., morepreferably 20 to 150° C. The reaction time is preferably 10 to 100hours, more preferably 1 to 20 hours.

(Step G)

Compound (8) is reacted with alkyl halide or aralkyl halide in thepresence of a base, to thereby synthesize compound (9).

The alkyl halide or aralkyl halide employable in this step, as well asreaction conditions, are the same as those described in step D ofSynthesis Method 1.

(Step H)

Compound (9) is reacted with an epoxidizing agent in a solvent in thepresence of a base, or with diazomethane in a solvent, to therebysynthesize compound (10).

In the former case, examples of the epoxidizing agents includetrimethylsulfoxonium iodide, trimethylsulfoxonium bromide. Preferably,the amount of the epoxidizing agent to be employed is 1 to 2 equivalentswith respect to compound (9). Examples of the bases include sodiumhydroxide, potassium hydroxide, barium hydroxide, sodium methoxide,sodium carbonate, potassium carbonate, and sodium hydride, with sodiumhydride being particularly preferred. Preferably, the amount of the baseto be employed is 1 to 5 equivalents with respect to compound (9). Asthe solvent, dimethyl sulfoxide, tetrahydrofuran, or a similar substancemay be used singly or as a mixture. The reaction temperature ispreferably −100° C. to the boiling point of the solvent, more preferably−40 to 50° C. The reaction time is preferably 5 minutes to 100 hours,more preferably 1 to 24 hours.

In the latter case using diazomethane, preferably, diethyl ether or asimilar substance is used as solvent. The reaction temperature ispreferably −50° C. to the boiling point of the solvent, more preferably−20 to 30° C. The reaction time is preferably 0.5 to 100 hours, morepreferably 1 to 24 hours.

(Step I)

Imidazole is reacted with compound (10), to thereby yield compound (1)of the present invention.

Preferably, the amount of the imidazole to be employed is 2 to 5equivalents with respect to compound (10). As the reaction solvent,N,N-dimethylformamide, dimethyl sulfoxide, methanol, ethanol,tetrahydrofuran, dioxane, or a similar substance may be used singly oras a mixture. In particular, use of N,N-dimethylformamide is preferred.Preferably, the reaction temperature is 20 to 100° C. The reaction timeis preferably 1 to 200 hours, more preferably 3 to 48 hours.

In the above-described synthesis methods, no particular limitation isimposed on the means for separating a target compound and the reactionsolvent. Examples of the means include, but are not limited to,filtration, extraction, distillation, wash, recrystallization, andchromatography.

The present compound (1) may be transformed into pharmaceuticallyacceptable salts; including inorganic salts based on inorganic acidssuch as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid,and hydrobromic acid; and organic salts based on organic acids such asfumaric acid, maleic acid, acetic acid, malic acid, tartaric acid,citric acid, methanesulfonic acid, and p-toluenesulfonic acid.

The compounds (1) of the present invention or salts thereof exhibitexcellent cytokine-like activity, in particular, G-CSF-like activity,and are endowed with high safety and high solubility to aqueous media,thus proving that they are suitably employed for formulation ofintravenous injections, and finding utility as a drug for prevention andtreatment of immune diseases accompanying reduced neutrophil count inanimals and humans.

The present compound (1) or their salts can be formed into variousdosage forms such as tablets, granules, powders, capsules, suspensions,injections, and suppositories, through a conventional method. When thecompound of the present invention is formed to have a solid form, avehicle may further be added, and when needed, other additives such as abinder, a disintegrant, a bulking agent, a coating agent, and a sugarcoating agent may also be added, and the resultant mixture is processedthrough a conventional method, to thereby produce solid dosage formssuch as tablets, granules, and capsules. In the case where injectionsare produced, the present compound is dissolved, dispersed, oremulsified in an aqueous carrier (e.g., distilled water for injection);or processed to a powder for preparing an injection, which powder isdissolved upon use. In the case where suppositories are prepared, anoily base or an emulsion base is added to the present compound, and theresultant mixture is processed through a conventional method.

The daily dose of the compound (1) of the present invention or saltsthereof for an adult is preferably about 0.05 mg to 5 g, more preferably1 to 100 mg, which is administered in a single dose or several divideddoses.

EXAMPLES

The present invention will next be described in more detail by way ofExamples, which should not be construed as limiting the invention.

Referential Example 1 Synthesis of2-Bromo-1-(5-chloro-2-hydroxyphenyl)-1-ethanone [Compound (4-1)]

A solution of 1-(5-chloro-2-hydroxyphenyl)-1-ethanone [compound (3-1)](41 g, 0.24 mol) in chloroform (250 mL) was added to a suspension ofcupric bromide (107.8 g, 0.48 mol) in ethyl acetate (250 mL), and theresultant mixture was stirred with reflux for 8 hours. After the mixturewas cooled on ice, dichloromethane (500 mL) was added, and insolublematter was filtered off, and the solvent was evaporated. Crystals thatprecipitated were collected by addition of dichloromethane, washed withdichloromethane, and dried with application of air flow, to therebyyield 2-bromo-1-(5-chloro-2-hydroxyphenyl)-1-ethanone as colorlesscrystals (35.8 g, yield: 60.0%).

¹H-NMR (CDCl₃) δ: 4.40 (2H, s), 6.97 (1H, d, J=8 Hz), 7.47 (1H, dd, J=8Hz, J=2 Hz), 7.73 (1H, d, J=2 Hz), 11.60 (1H, s).

Referential Example 2 Synthesis of2-Bromo-1-(5-fluoro-2-hydroxyphenyl)-1-ethanone [Compound (4-2)]

The general procedure of Referential Example 1 was repeated, except that1-(5-fluoro-2-hydroxyphenyl)-1-ethanone [compound (3-2)] was usedinstead of 1-(5-chloro-2-hydroxyphenyl)-1-ethanone [compound (3-1)], tothereby yield 2-bromo-1-(5-fluoro-2-hydroxyphenyl)-1-ethanone ascolorless crystals (yield: 80.4%).

¹H-NMR (CDCl₃) δ: 4.38 (2H, s), 6.8-7.6 (3H, m), 11.47 (1H, s).

Referential Example 3 Synthesis of1-(5-Chloro-2-hydroxyphenyl)-2-(1H-1-imidazolyl)-1-ethanone [compound(5-1)]

Imidazole (17.9 g, 0.26 mol) was added to a solution of2-bromo-1-(5-chloro-2-hydroxyphenyl)-1-ethanone [compound (4-1)] (21.8g, 87.7 mmol) in N,N-dimethylformamide (50 mL), and the resultantmixture was stirred for 17 hours at room temperature. The reactionmixture was added to ice-water, followed by extraction with ethylacetate. The organic layer was washed with water, and dried overmagnesium sulfate. Subsequently, the solvent was removed under reducedpressure. The residue was purified through silica gel columnchromatography, followed by drying under reduced pressure, to therebyyield 1-(5-chloro-2-hydroxyphenyl)-2-(1H-1-imidazolyl)-1-ethanone ascolorless crystals (17.1 g, yield: 82.4%).

¹H-NMR (DMSO-d₆) δ: 5.56 (2H, s), 6.7-7.9 (6H, m), 10.3-10.7 (1H, m).

Referential Example 4 Synthesis of1-(5-Fluoro-2-hydroxyphenyl)-2-(1H-1-imidazolyl)-1-ethanone [Compound(5-2)]

The general procedure of Referential Example 3 was repeated, except that2-bromo-1-(5-fluoro-2-hydroxyphenyl)-1-ethanone [compound (4-2)], tothereby yield1-(5-fluoro-2-hydroxyphenyl)-2-(1H-1-imidazolyl)-1-ethanone as colorlesscrystals (yield: 86.5%).

¹H-NMR (DMSO-d₆) δ: 5.81 (2H, s), 6.7-8.0 (7H, m).

Referential Example 5 Synthesis of 4-Chlorophenyl Acetate [Compound(7-1)]

Acetic anhydride (80.1 g, 0.78 mol) and one drop of concentratedsulfuric acid were added to 4-chlorophenol [compound (6-1)] (100 g, 0.78mol) at room temperature, and the mixture was stirred for 17 hours. Theresultant mixture was added to ice-water, and converted to alkaline bythe addition of sodium hydroxide (20% aqueous solution), followed byextraction with diethyl ether. The organic layer was washed with water,dried over magnesium sulfate. Subsequently, the solvent was removedunder reduced pressure, and then the residue was subjected todistillation under reduced pressure (81° C./1.5 mmHg), to thereby yield4-chlorophenyl acetate as a colorless oily product (130 g, yield:98.0%).

¹H-NMR (CDCl₃) δ: 2.25 (3H, s), 6.97 (2H, d, J=8 Hz), 7.30 (2H, d, J=8Hz).

Referential Example 6 Synthesis of1-(5-Chloro-2-hydroxyphenyl)-1-ethanone [Compound (3-1)]

4-chlorophenyl acetate [compound (7-1)] (50 g, 0.293 mol) and aluminumchloride powder (117 g, 0.88 mol) were mixed, and the mixture wasstirred for 2 hours at 110° C. The resultant mixture was cooled to roomtemperature and added to ice-water, followed by extraction with diethylether. The organic layer was washed with water, and dried over magnesiumsulfate. The solvent was removed under reduced pressure, to therebyyield 1-(5-chloro-2-hydroxyphenyl)-1-ethanone as colorless crystals(40.2 g, yield: 80.4%).

¹H-NMR (CDCl₃) δ: 2.62 (3H, s), 6.90 (1H, d, J=8 Hz), 7.39 (1H, dd, J=8Hz, J=2 Hz), 7.65 (1H, d, J=2 Hz), 12.14 (1H, s).

Referential Example 7 Synthesis of1-(2-(Benzyloxy)-5-chlorophenyl)-1-ethanone [Compound (9-1)]

Potassium carbonate powder (2.9 g, 21.0 mmol) and benzyl bromide (2.3 g,18.2 mmol) was added to a solution of1-(5-chloro-2-hydroxyphenyl)-1-ethanone [compound (8-1)] (3.0 g, 17.6mmol) in dimethylformamide (50 mL), and the mixture was stirred for 3hours at 60° C. After cooling, ice-water was added to the resultantmixture, followed by extraction with ethyl acetate. The mixture waswashed with aqueous sodium hydroxide (5%) and water, and dried overmagnesium sulfate. The solvent was removed under reduced pressure, tothereby yield compound (9-1) as a colorless oily product (4.6 g, yield:99.9%).

¹H-NMR (CDCl₃) δ: 2.58 (3H, s), 5.14 (2H, s), 6.8-8.8 (3H, m), 7.39 (5H,s).

Referential Example 8 Synthesis of2-(2-(Benzyloxy)-5-chlorophenyl)-2-methyloxirane [Compound (10-1)]

A suspension of 60% sodium hydride (1.2 g,30 mmol) in tetrahydrofuran(30 mL)-dimethyl sulfoxide (30 mL) was heated at external temperature of60° C., and trimethylsulfoxonium iodide (6.5 g, 30 mmol) was addedportionwise to the mixture. The resultant mixture was stirred for 1 hourand then cooled to room temperature. A solution of1-(2-(benzyloxy)-5-chlorophenyl)-1-ethanone [compound (9-1)] (5.0 g, 19mmol) in tetrahydrofuran (10 mL) was added dropwise to the mixture, andthe mixture was stirred for 3 hours at external temperature of 60° C.After the mixture was cooled to room temperature, the mixture was addedto ice-water, followed by extraction with diethyl ether. The extractedsubstance was washed with water and dried over magnesium sulfate. Thesolvent was removed under reduced pressure, to thereby yield2-(2-(benzyloxy)-5-chlorophenyl)-2-methyloxirane as a colorless oilyproduct (yield: 4.4 g). The obtained product was used in the subsequentreaction step without purification.

¹H-NMR (CDCl₃) δ: 1.63 (3H, s), 2.74 (1H, d, J=5.3 Hz), 2.90 (1H, d,J=5.3 Hz), 5.10 (2H, s), 6.6-7.6 (3H, m), 7.38 (5H, s).

Example 1 Synthesis of4-Chloro-2-(1-hydroxy-2-(1H-1-imidazolyl)-1-methylethyl)phenol [Compound(2-1)]

A suspension of1-(5-chloro-2-hydroxyphenyl)-2-(1H-1-imidazolyl)-1-ethanone [compound(5-1)] (2.37 g, 10 mmol) in tetrahydrofuran (200 mL) was added to asolution of methylmagnesium bromide in tetrahydrofuran (1 mol/L, 100 mL,0.1 mol) under cooling with ice, and the mixture was stirred for 17hours at room temperature. The solvent was removed, a solution ofsaturated ammonium chloride was added to the resultant mixture, followedby extraction with ethyl acetate. The organic layer was washed withwater and dried over magnesium sulfate. The solvent was removed underreduced pressure. Subsequently, the residue was purified through silicagel column chromatography and dried under reduced pressure, to therebyyield 4-chloro-2-(1-hydroxy-2-(1H-1-imidazolyl)-1-methylethyl)phenol ascolorless crystals (620 mg, yield: 24.5%).

¹H-NMR (CDCl₃) δ: 1.63 (3H, s), 4.13 (1H, d, J=14 Hz), 4.32 (1H, d, J=14Hz), 6.6-7.3 (5H, m), 7.37 (1H, s), 8.90 (2H, br.).

Example 2 Synthesis of4-Chloro-2-(1-hydroxy-1-(1H-1-imidazolylmethyl)butyl)phenol [Compound(2-2)]

The general procedure of Example 1 was repeated using1-(5-chloro-2-hydroxyphenyl)-2-(1H-1-imidazolyl)-1-ethanone [compound(5-1)] and n-propylmagnesium bromide, to thereby yield4-chloro-2-(1-hydroxy-1-(1H-1-imidazolylmethyl)butyl)phenol as acolorless oily product (yield: 50.4%).

¹H-NMR (CDCl₃) δ: 0.7-1.1 (3H, m), 1.1-2.4 (4H, m), 4.14 (1H, d, J=14Hz), 4.28 (1H, d, J=14 Hz), 6.5-7.5 (6H, m), 7.7-8.1 (2H, m).

Example 3 Synthesis of4-Chloro-2-(1-cyclopropyl-1-hydroxy-2-(1H-1-imidazolyl)ethyl)phenol[Compound (2-3)]

The general procedure of Example 1 was repeated using1-(5-chloro-2-hydroxyphenyl)-2-(1H-1-imidazolyl)-1-ethanone [compound(5-1)] and cyclopropylmagnesium bromide, to thereby yield4-chloro-2-(1-cyclopropyl-1-hydroxy-2-(1H-1-imidazolyl)ethyl)phenol ascolorless crystals (yield: 36.0%).

¹H-NMR (CDCl₃) δ: 0.2-0.7 (4H, m), 1.0-1.5 (1H, m), 4.30 (1H, d, J=14Hz), 4.38 (1H, d, J=14 Hz), 6.6-7.5 (6H, m), 7.9-8.3 (2H, br.).

Example 4 Synthesis of4-Fluoro-2-(1-hydroxy-2-(1H-1-imidazolyl)-1-methylethyl)phenol [Compound(2-4)]

The general procedure of Example 1 was repeated using1-(5-fluoro-2-hydroxyphenyl)-2-(1H-1-imidazolyl)-1-ethanone [compound(5-2)] and methylmagnesium bromide, to thereby yield4-fluoro-2-(1-hydroxy-2-(1H-1-imidazolyl)-1-methylethyl)phenol ascolorless crystals (yield: 53.2%).

¹H-NMR (CDCl₃) δ: 1.57 (3H, s), 4.12 (1H, d, J=14 Hz), 4.27 (1H, d, J=14Hz), 6.6-6.9 (5H, m), 7.35 (1H, s), 8.23 (2H, br.).

Example 5 Synthesis of4-Fluoro-2-(1-hydroxy-1-(1H-1-imidazolylmethyl)butyl)phenol [Compound(2-5)]

The general procedure of Example 1 was repeated using1-(5-fluoro-2-hydroxyphenyl)-2-(1H-1-imidazolyl)-1-ethanone [compound(5-2)] and n-propylmagnesium bromide, to thereby yield4-fluoro-2-(1-hydroxy-1-(1H-1-imidazolylmethyl)butyl)phenol as acolorless oily product (yield: 31.3%).

¹H-NMR (CDCl₃) δ: 0.7-1.1 (3H, m), 1.1-2.3 (4H, m), 4.18 (1H, d, J=14Hz), 4.29 (1H, d, J=14 Hz), 6.5-7.5 (6H, m), 8.1-8.8 (2H, m).

Example 6 Synthesis of2-(5-Chloro-2-(n-nonyloxy)phenyl)-1-(1H-1-imidazolyl)-2-propanol[Compound (1-1)]

Sodium hydroxide powder (20 mg, 0.5 mmol) and n-nonyl bromide (122 mg,0.59 mmol) were added to a solution of4-chloro-2-(1-hydroxy-2-(1H-1-imidazolyl)-1-methylethyl)phenol [compound(2-1)] (100 mg, 0.40 mmol) in N,N-dimethylformamide (1 mL), and themixture was stirred for 17 hours at room temperature. Ice-water wasadded to the resultant mixture, followed by extraction with ethylacetate. The organic layer was washed with water and dried overmagnesium sulfate. The solvent was removed under reduced pressure. Theresidue was purified through silica gel column chromatography and driedunder reduced pressure, to thereby yield2-(5-chloro-2-(n-nonyloxy)phenyl)-1-(1H-1-imidazolyl)-2-propanol as acolorless oily product (81.7 mg, yield: 54.5%).

¹H-NMR (CDCl₃) δ: 0.7-1.0 (3H, m), 1.0-2.2 (14H, m), 1.53 (3H, s), 4.02(2H, t, J=6 Hz), 4.33 (2H, s), 4.83 (1H, s), 6.6-7.6 (6H, m).

Example 7 Synthesis of2-(5-Chloro-2-(n-heptyloxy)phenyl)-1-(1H-1-imidazolyl)-2-propanol[Compound (1-2)]

The general procedure of Example 6 was repeated using4-chloro-2-(1-hydroxy-2-(1H-1-imidazolyl)-1-methylethyl)phenol [compound(2-1)] and n-heptyl bromide, to thereby yield2-(5-chloro-2-(n-heptyloxy)phenyl)-1-(1H-1-imidazolyl)-2-propanol as acolorless oily product (yield: 74.3%).

¹H-NMR (CDCl₃) δ: 0.7-1.0 (3H, m), 1.0-2.2 (10H, m), 1.53 (3H, s), 4.02(2H, t, J=6 Hz), 4.33 (2H, s), 4.83 (1H, s), 6.6-7.6 (6H, m).

Example 8 Synthesis of2-(5-Chloro-2-(n-octyloxy)phenyl)-1-(1H-1-imidazolyl)-2-propanol[Compound (1-3)]

The general procedure of Example 6 was repeated using4-chloro-2-(1-hydroxy-2-(1H-1-imidazolyl)-1-methylethyl)phenol [compound(2-1)] and n-octyl bromide, to thereby yield2-(5-chloro-2-(n-octyloxy)phenyl)-1-(1H-1-imidazolyl)-2-propanol as acolorless oily product (yield: 75.5%).

¹H-NMR (CDCl₃) δ: 0.7-1.0 (3H, m), 1.0-2.2 (12H, m), 1.53 (3H, s), 4.02(2H, t, J=6 Hz), 4.31 (2H, s), 4.79 (1H, s), 6.6-7.6 (6H, m).

Example 9 Synthesis of2-(5-Chloro-2-((2,4-dichlorobenzyl)oxy)phenyl)-1-(1H-1-imidazolyl)-2-propanol[Compound (1-4)]

The general procedure of Example 6 was repeated using4-chloro-2-(1-hydroxy-2-(1H-1-imidazolyl)-1-methylethyl)phenol [compound(2-1)] and 2,4-dichlorobenzyl chloride, to thereby yield2-(5-chloro-2-((2,4-dichlorobenzyl)oxy)phenyl)-1-(1H-1-imidazolyl)-2-propanolas colorless crystals (yield: 51.9%).

¹H-NMR (CDCl₃) δ: 1.49 (3H, s), 4.29 (2H, s), 4.80 (1H, s), 5.16 (2H,s), 6.6-7.7 (9H, m).

Example 10 Synthesis of2-(5-Chloro-2-(phenethyloxy)phenyl)-1-(1H-1-imidazolyl)-2-propanol[Compound (1-5)]

The general procedure of Example 6 was repeated using4-chloro-2-(1-hydroxy-2-(1H-1-imidazolyl)-1-methylethyl)phenol [compound(2-1)] and phenethyl chloride, to thereby yield2-(5-chloro-2-(phenethyloxy)phenyl)-1-(1H-1-imidazolyl)-2-propanol as acolorless oily product (yield: 12.4%).

¹H-NMR (CDCl₃) δ: 1.38 (3H, s), 3.14 (2H, t, J=6 Hz), 4.06 (2H, s), 4.30(2H, t, J=6 Hz), 4.43 (1H, s), 6.4-7.4 (6H, m), 7.29 (5H, s).

Example 11 Synthesis of2-(5-Chloro-2-(n-pentyloxy)phenyl)-1-(1H-1-imidazolyl)-2-pentanol[Compound (1-6)]

The general procedure of Example 6 was repeated using4-chloro-2-(1-hydroxy-1-(1H-1-imidazolylmethyl)butyl)phenol [compound(2-2)] and n-pentyl bromide, to thereby yield2-(5-chloro-2-(n-pentyloxy)phenyl)-1-(1H-1-imidazolyl)-2-pentanol as acolorless oily product (yield: 79.9%).

¹H-NMR (CDCl₃) δ: 0.6-1.1 (6H, m), 1.1-2.5 (10H, m), 4.00 (2H, t, J=6Hz), 4.22 (1H, d, J=14 Hz), 4.34 (1H, d, J=14 Hz), 4.5-4.7 (1H, br.),6.5-7.6 (6H, m).

Example 12 Synthesis of2-(5-Chloro-2-((2,4-dichlorobenzyl)oxy)phenyl)-1-(1H-1-imidazolyl)-2-pentanol[Compound (1-7)]

The general procedure of Example 6 was repeated using4-chloro-2-(1-hydroxy-1-(1H-1-imidazolylmethyl)butyl)phenol [compound(2-2)] and 2,4-dichlorobenzyl chloride, to thereby yield2-(5-chloro-2-((2,4-dichlorobenzyl)oxy)phenyl)-1-(1H-1-imidazolyl)-2-pentanolas a colorless oily product (yield: 30.0%).

¹H-NMR (CDCl₃) δ: 0.6-1.0 (3H, m), 1.0-2.5 (4H, m), 4.20 (1H, d, J=14Hz), 4.31 (1H, d, J=14 Hz), 4.3-4.6 (1H, br.), 5.11 (2H, s), 6.4-7.6(9H, m).

Example 13 Synthesis of2-(5-Chloro-2-((2-fluorophenethyl)oxy)phenyl)-1-(1H-1-imidazolyl)-2-pentanol[Compound (1-8)]

The general procedure of Example 6 was repeated using4-chloro-2-(1-hydroxy-1-(1H-1-imidazolylmethyl)butyl)phenol [compound(2-2)] and 2-fluorophenethyl chloride, to thereby yield2-(5-chloro-2-((2-fluorophenethyl)oxy)phenyl)-1-(1H-1-imidazolyl)-2-pentanolas a colorless oily product (yield: 7.4%).

¹H-NMR (CDCl₃) δ: 0.6-1.0 (3H, m), 1.0-2.3 (4H, m), 3.20 (2H, t, J=6Hz), 4.1-4.5 (5H, m), 6.5-7.6 (10H, m).

Example 14 Synthesis of1-(5-Chloro-2-(n-pentyloxy)phenyl)-1-cyclopropyl-2-(1H-1-imidazolyl)-1-ethanol[Compound (1-9)]

The general procedure of Example 6 was repeated using4-chloro-2-(1-cyclopropyl-1-hydroxy-2-(1H-1-imidazolyl)ethyl)phenol[compound (2-3)] and n-pentyl bromide, to thereby yield1-(5-chloro-2-(n-pentyloxy)phenyl)-1-cyclopropyl-2-(1H-1-imidazolyl)-1-ethanolas a colorless oily product (yield: 71.3%).

¹H-NMR (CDCl₃) δ: 0.1-0.7 (4H, m), 0.7-1.1 (3H, m), 1.1-2.3 (7H, m),4.02 (2H, t, J 6 Hz), 4.20 (1H, s), 4.36 (1H, d, J=14 Hz), 4.52 (1H, d,J=14 Hz), 6.6-7.5 (6H, m).

Example 15 Synthesis of1-(5-Chloro-2-(n-octyloxy)phenyl)-1-cyclopropyl-2-(1H-1-imidazolyl)-1-ethanol[Compound (1-10)]

The general procedure of Example 6 was repeated using4-chloro-2-(1-cyclopropyl-1-hydroxy-2-(1H-1-imidazolyl)ethyl)phenol[compound (2-3)] and n-octyl bromide, to thereby yield1-(5-chloro-2-(n-octyloxy)phenyl)-1-cyclopropyl-2-(1H-1-imidazolyl)-1-ethanolas a colorless oily product (yield: 58.1%).

¹H-NMR (CDCl₃) δ: 0.1-0.7 (4H, m), 0.7-1.1 (3H, m), 1.1-2.3 (13H, m),4.02 (2H, t, J=6 Hz), 4.34 (1H, d, J=14 Hz), 4.52 (1H, d, J=14 Hz),6.6-7.5 (6H, m).

Example 16 Synthesis of1-(5-chloro-2-(phenethyloxy)phenyl)-1-cyclopropyl-2-(1H-1-imidazolyl)-1-ethanol[Compound (1-11)]

The general procedure of Example 6 was repeated using4-chloro-2-(1-cyclopropyl-1-hydroxy-2-(1H-1-imidazolyl)ethyl)phenol[compound (2-3)] and phenethyl chloride, to thereby yield1-(5-chloro-2-(phenethyloxy)phenyl)-1-cyclopropyl-2-(1H-1-imidazolyl)-1-ethanolas colorless crystals (yield: 26.4%).

¹H-NMR (CDCl₃) δ: 0.1-0.6 (4H, m), 1.0-1.5 (1H, m), 3.16 (2H, t, J=6Hz), 4.14 (1H, d, J=14 Hz), 4.30 (2H, t, J=6 Hz), 4.34 (1H, d, J=14 Hz),4.47 (1H, s), 6.5-7.5 (6H, m), 7.28 (5H, s).

Example 17 Synthesis of2-(5-Fluoro-2-(n-heptyloxy)phenyl)-1-(1H-1-imidazolyl)-2-propanol[Compound (1-12)]

The general procedure of Example 6 was repeated using4-fluoro-2-(1-hydroxy-2-(1H-1-imidazolyl)-1-methylethyl)phenol [compound(2-4)] and n-heptyl bromide, to thereby yield2-(5-fluoro-2-(n-heptyloxy)phenyl)-1-(1H-1-imidazolyl)-2-propanol ascolorless crystals (yield: 94.3%).

¹H-NMR (CDCl₃) δ: 0.7-1.0 (3H, m), 1.0-2.2 (10H, m), 1.53 (3H, s), 4.02(2H, t, J=6 Hz), 4.33 (2H, s), 5.38(1H, s), 6.6-7.4 (6H, m).

Example 18 Synthesis of2-(5-Fluoro-2-(n-octyloxy)phenyl)-1-(1H-1-imidazolyl)-2-propanol[Compound (1-13)]

The general procedure of Example 6 was repeated using4-fluoro-2-(1-hydroxy-2-(1H-1-imidazolyl)-1-methylethyl)phenol [compound(2-4)] and n-octyl bromide, to thereby yield2-(5-fluoro-2-(n-octyloxy)phenyl)-1-(1H-1-imidazolyl)-2-propanol as acolorless oily product (yield: 86.1%).

¹H-NMR (CDCl₃) δ: 0.7-1.0 (3H, m), 1.0-2.2 (12H, m), 1.53 (3H, s), 4.00(2H, t, J=6 Hz), 4.35 (2H, s), 5.47 (1H, s), 6.6-7.4 (6H, m).

Example 19 Synthesis of2-(5-Fluoro-2-(n-nonyloxy)phenyl)-1-(1H-1-imidazolyl)-2-propanol[Compound (1-14)]

The general procedure of Example 6 was repeated using4-fluoro-2-(1-hydroxy-2-(1H-1-imidazolyl)-1-methylethyl)phenol [compound(2-4)] and n-nonyl bromide, to thereby yield2-(5-fluoro-2-(n-nonyloxy)phenyl)-1-(1H-1-imidazolyl)-2-propanol as acolorless oily product (yield: 88.4%).

¹H-NMR (CDCl₃) δ: 0.7-1.0 (3H, m), 1.0-2.2 (14H, m), 1.53 (3H, s), 4.00(2H, t, J=6 Hz), 4.33 (2H, s), 4.84 (1H, s), 6.6-7.4 (6H, m).

Example 20 Synthesis of2-(2-((2,4-Dichlorobenzyl)oxy)-5-fluorophenyl)-1-(1H-1-imidazolyl)-2-propanol[Compound (1-15)]

The general procedure of Example 6 was repeated using4-fluoro-2-(1-hydroxy-2-(1H-1-imidazolyl)-1-methylethyl)phenol [compound(2-4)] and 2,4-dichlorobenzyl chloride, to thereby yield2-(2-((2,4-dichlorobenzyl)oxy)-5-fluorophenyl)-1-(1H-1-imidazolyl)-2-propanolas colorless crystals (yield: 31.6%).

¹H-NMR (CDCl₃) δ: 1.50 (3H, s), 4.24 (1H, s), 4.32 (2H, s), 5.19 (2H,s), 6.6-7.6 (9H, m).

Example 21 Synthesis of2-(5-Fluoro-2-(phenethyloxy)phenyl)-1-(1H-1-imidazolyl)-2-propanol[Compound (1-16)]

The general procedure of Example 6 was repeated using4-fluoro-2-(1-hydroxy-2-(1H-1-imidazolyl)-1-methylethyl)phenol [compound(2-4)] and phenethyl chloride, to thereby yield2-(5-fluoro-2-(phenyloxy)phenyl)-1-(1H-1-imidazolyl)-2-propanol as acolorless oily product (yield: 13.4%).

¹H-NMR (CDCl₃) δ: 1.41 (3H, s), 3.17 (2H, t, J=6 Hz), 4.07 (2H, s), 4.30(2H, t, J=6 Hz), 4.57 (1H, s), 6.5-7.5 (6H, m), 7.29 (5H, s).

Example 22 Synthesis of2-(5-Fluoro-2-(n-pentyloxy)phenyl)-1-(1H-1-imidazolyl)-2-pentanol[Compound (1-17)]

The general procedure of Example 6 was repeated using4-fluoro-2-(1-hydroxy-1-(1H-1-imidazolylmethyl)butyl)phenol [compound(2-5)] and n-pentyl bromide, to thereby yield2-(5-fluoro-2-(n-pentyloxy)phenyl)-1-(1H-1-imidazolyl)-2-pentanol as acolorless oily product (yield: 80.3%).

¹H-NMR (CDCl₃) δ: 0.6-1.0 (3H, m), 0.94 (3H, t, J=6 Hz), 1.0-2.5 (10H,m), 3.99 (2H, t, J=6 Hz), 4.14 (1H, s), 4.29 (1H, d, J=14 Hz), 4.42 (1H,d, J=14 Hz), 6.5-7.5 (6H, m).

Example 23 Synthesis of2-(2-((4-Chlorobenzyl)oxy)-5-fluorophenyl)-1-(1H-1-imidazolyl)-2-pentanol[Compound (1-18)]

The general procedure of Example 6 was repeated using4-fluoro-2-(1-hydroxy-1-(1H-1-imidazolylmethyl)butyl)phenol [compound(2-5)] and 4-chlorobenzyl chloride, to thereby yield2-(2-((4-chlorobenzyl)oxy)-5-fluorophenyl)-1-(1H-1-imidazolyl)-2-pentanolas a colorless oily product (yield: 8.1%).

¹H-NMR (CDCl₃) δ: 0.6-1.0 (3H, m), 1.0-2.5 (4H, m), 3.2-3.5 (1H, m),4.25 (1H, d, J=14 Hz), 4.34 (1H, d, J=14 Hz), 5.02 (2H, s), 6.6-7.6(10H, m).

Example 24 Synthesis of2-(2-((2,4-Dichlorobenzyl)oxy)-5-fluorophenyl)-1-(1H-1-imidazolyl)-2-pentanol[Compound (1-19)]

The general procedure of Example 6 was repeated using4-fluoro-2-(1-hydroxy-1-(1H-1-imidazolylmethyl)butyl)phenol [compound(2-5)] and 2,4-dichlorobenzyl chloride, to thereby yield2-(2-((2,4-dichlorobenzyl)oxy)-5-fluorophenyl)-1-(1H-1-imidazolyl)-2-pentanolas a colorless oily product (yield: 13.0%).

¹H-NMR (CDCl₃) δ: 0.6-1.0 (3H, m), 1.0-2.5 (4H, m), 3.2-3.6 (1H, m),4.25 (1H, d, J=14 Hz), 4.36 (1H, d, J=14 Hz), 5.15 (2H, s), 6.5-7.6 (9H,m).

Example 25 Synthesis of2-(5-Fluoro-2-((2-fluorophenethyl)oxy)phenyl)-1-(1H-1-imidazolyl)-2-pentanol[Compound (1-20)]

The procedure of Example 6 was repeated using4-fluoro-2-(1-hydroxy-1-(1H-1-imidazolylmethyl)butyl)phenol [compound(2-5)] and 2,4-dichlorobenzyl chloride, to thereby yield2-(5-fluoro-2-((2-fluorophenethyl)oxy)phenyl)-1-(1H-1-imidazolyl)-2-pentanolas a colorless oily product (yield: 8.0%).

¹H-NMR (CDCl₃) δ: 0.6-1.0 (3H, m), 1.0-2.3 (4H, m), 3.20 (2H, t, J=6Hz), 3.8-4.5 (5H, m), 6.5-7.5 (10H, m).

Example 26 Synthesis of2-(2-(Benzyloxy)-5-chlorophenyl)-1-(1H-1-imidazolyl)-2-propanol[Compound (1-21)]

Imidazole (4.0 g, 58.8 mmol) and potassium t-butoxide (6.5 g, 57.9 mmol)were added to a solution of2-(2-(benzyloxy)-5-chlorophenyl)-2-methyloxirane [compound (10-1),obtained in Referential Example 8] (4.4 g) in dimethylformamide (60 mL),and the mixture was stirred for 12 hours at 80° C. After the mixture wascooled to room temperature, ice-water was added thereto, followed byextraction with ethyl acetate. The organic layer was washed with waterand dried over magnesium sulfate. The solvent was removed under reducedpressure. The residue was purified through silica gel columnchromatography and dried under reduced pressure, to thereby yield2-(2-(benzyloxy)-5-chlorophenyl)-1-(1H-1-imidazolyl)-2-propanol ascolorless crystals (5.0 g, yield: 75.8%).

¹H-NMR (CDCl₃) δ: 1.47 (3H, s), 4.27 (2H, s), 5.09 (2H, s), 6.5-7.6 (6H,m), 7.42 (5H, s). MS (FAB): 345, 343 (M+H).

Test Example 1 Creation of a Human G-CSF-dependent Cell Strain (invitro)

Cloning of human G-CSF receptor gene was performed through RT-PCR byusing primers created on the basis of the human G-CSF receptor geneusing a human spleen cDNA library. The thus cloned human G-CSF receptorgene inserted to a multicloning site of an expression vector.Subsequently, the gene-inserted expression vector was transformed intoE. coli for multiplication, and the expression vector was extracted. Analiquot of the expression vector (20 μg) was added to host cells(BAF/B03) prepared to 5×10⁷ cells/800 μL K-PBS (30.8 mM NaCl, 120.7 mMKCl, 8.1 mM Na₂HPO₄, 1.46 mM KH₂PO₄, 5 mM MgCl₂), followed by standingstill at 4° C. for 15 minutes. For gene transfer, electroporation wasperformed at a voltage of 280 V and a capacitance of 950 μF. Aftercompletion of gene transfer, the cells were suspended in a DMEM medium((+) 10% FCS) containing recombinant human G-CSF (rhG-CSF, 20 ng/mL) andpuromycin (2 μg/mL), whereby a human G-CSF dependent cell strain wascreated.

For studying the cell multiplication ability with respect to rhG-CSF,the following procedure was carried out. Briefly, cells in which thehuman G-CSF receptor gene had been inserted (BAF/hGCSFR) were collected,and washed with PBS(−). Subsequently, the cells were suspended in DMEMmedium, and the medium containing the cells was prepared to the wells ofa 96-well plate (5×10⁴ cells/well). Then, rhG-CSF was added to the wells(0, 0.006, 0.025, 0.1, 0.4 ng/mL), and the cells in the wells werecultured under 5% CO₂ for 48 hours at 37° C. After completion ofincubation, a solution of WST-1/1-methoxy PMS was added to the wells(final concentration: 5 mM), and the cells in the wells were cultured ina CO₂ incubator for 2 hours. After completion of incubation, theabsorbance of the cultured solution was measured at a wavelength of 450nm. The results are shown in FIG. 1 (mean±standard error, n=5).

Test Example 2 Retrieval of Compounds Inducing Cell Growth of a HumanG-CSF Dependent Cell Strain (in vitro)

A solution prepared by diluting a compound with DMEM was added to thewells of a 96-well microplate (10 μL/well). The human G-CSF-dependentcell strain was added to the wells (5.0×10⁴ cells/100 μL), followed byincubation for 48 hours at 37° C. After completion of incubation, asolution of WST-1/1-methoxy PMS (final concentration: 5 mM) was added tothe wells, followed by incubation in a CO₂ incubator for 2 hours. Aftercompletion of incubation, the absorbance was measured at a wavelength of450 nm. The results are shown in FIG. 2 (mean±standard error, n=5).

Test Example 3 Human-G-CSF Equivalent Concentrations Corresponding toRespective Compound Concentrations

Using the test data obtained from Test Example 1, human-G-CSF equivalentconcentrations (ng/mL) corresponding to respective compoundconcentrations employed in Test Example 2 were calculated. The resultsare shown in Table 1 (mean±standard error, n=5).

TABLE 1 compound conc. compound 10⁻⁶ M 10⁻⁷ M 10⁻⁸ M (1-1) 0.131 ±0.001  0.032 ± 0.0003 0.015 ± 0.0001 (1-2) 0.049 ± 0.0004 0.037 ± 0.00030.013 ± 0.0001 (1-3) 0.057 ± 0.0005 0.057 ± 0.0005 0.018 ± 0.0001

Industrial Applicability

The compounds (1) or salts thereof according to the present inventionexhibit excellent cytokine-like activity, in particular, G-CSF-likeactivity, and are endowed with high safety and high solubility toaqueous media, thus proving that they are suitably employed forformulation of intravenous injections, and finding utility as a drug forprevention and treatment of immune diseases accompanying reducedneutrophil count in animals and humans.

What is claimed is:
 1. An imidazole derivative represented by thefollowing formula (1):

wherein R¹ represents a lower alkyl group, R² represents an at least aC₆ alkyl group, or an aralkyl group which may optionally behalo-substituted, and X represents a halogen atom, or a salt or ahydrate thereof.
 2. An imidazole derivative represented by the followingformula (2):

wherein R¹ represents a lower alkyl group, and X¹ represents a halogenatom, or a salt thereof.
 3. The imidazole derivative of claim 1, whereinR¹ is linear alkyl group.
 4. The imidazole derivative of claim 1,wherein R¹ is branched alkyl group.
 5. The imidazole derivative of claim1, wherein R¹ is cyclic alkyl group.
 6. The imidazole derivative ofclaim 1, wherein R² is n-hexyl, n-heptyl or n-octyl.
 7. The imidazolederivative of claim 1, wherein R² is n-nonyl or n-decyl.
 8. Theimidazole derivative of claim 1, wherein R² is n-undecyl or n-dodecyl.9. The imidazole derivative of claim 1, wherein R² is a linear alkylgroup.
 10. The imidazole derivative of claim 1, wherein R² is a branchedalkyl group.
 11. The imidazole derivative of claim 1, wherein R² isphenyl C₁-C₅ alkyl.
 12. The imidazole derivative of claim 1, wherein R²is a halogen-substituted aralkyl group.
 13. The imidazole derivative ofclaim 1, wherein X¹ is a fluorine atom.
 14. The imidazole derivative ofclaim 1, wherein X¹ is a chlorine atom.
 15. A salt of the imidazolederivative of claim 1 selected from the group consisting of thehydrochloride, nitrate, hydrobromide, p-toluenesulfonate,methanesulfonate, fumate, succinate and lactate.
 16. An isomer ormixture of isomers of the derivative of claim
 1. 17. A compositioncomprising the derivative of claim 1 or a salt thereof, and apharmaceutically acceptable carrier.
 18. The composition of claim 17 ina solid dosage form.
 19. The composition of claim 17 in a dissolved,dispersed or emulsified form.
 20. The composition of claim 17 in aninjectable form.
 21. The composition of claim 17 in the form of atablet, capsule or granule.
 22. A method for manufacturing a drug or atherapeutic composition comprising: admixing the imidazole derivative ofclaim 1 or a salt or a hydrate thereof with a pharmaceuticallyacceptable carrier or excipient.
 23. A method for inducing G-CSF-likeactivity, comprising administering to a subject in need thereof theimidazole derivative of claim
 1. 24. The method of claim 23 comprisingadministering a dose ranging from 0.05 mg to 5 g of said imidazolederivative.
 25. A method for treating an immune disease accompanying apathological condition with a reduction in neutrophils, comprising:administering to a subject in need thereof the imidazole derivative ofclaim 1 or a salt thereof.
 26. A method for making the imidazolederivative of claim 1 comprising: reacting an imidazole derivative offormula (2) with an alkyl halide or aralkyl halide in the presence of abase, wherein formula (2) is:

wherein R¹ represents a lower alkyl group, and X¹ represents a halogenatom, or a salt thereof.
 27. The imidazole derivative of claim 2,wherein R¹ is a linear alkyl group.
 28. The imidazole derivative ofclaim 2, wherein R¹ is a branched alkyl group.
 29. The imidazolederivative of claim 2, wherein R¹ is cyclic alkyl group.
 30. Theimidazole derivative of claim 2, wherein X¹ is a fluorine atom.
 31. Theimidazole derivative of claim 2, wherein X¹ is a chlorine atom.